​DR. TOM ROBINSON

Minimal cells created, manipulated and analysed using microfluidic systems

Giant unilamellar vesicles (GUVs) are now considered the gold standard for the bottom-up construction of synthetic minimal cells. However, traditional techniques lack the ability to encapsulate the required machinery or to control their sizes. Microfluidic systems, on the other hand, are ideally suited to this challenging task (Robinson. Advanced Biosystems, 2019). First, we present a microfluidic platform based on droplet technology which is able to produce monodisperse biomimetic GUVs with high encapsulation - ideal for synthetic cell construction. Microfluidics is also used to produce artificial minimal cells with multiple membrane compartments, either by encapsulation of smaller vesicles or by complete on-chip methods. Next, we present novel methods for the capture and analysis of artificial cells (Robinson et al., Biomicrofluidics 2013; Yandralli & Robinson, Lab on a Chip 2019) including a new high-throughput device able to perform up to 8 separate experiments with 800 GUVs each as well as an integrated platform able to produce and capture GUVs in the same device (Yandralli & Robinson, submitted). Finally, we present two applications of our multi-compartment minimal cells. The first system we designed to exhibit self-organisation of its internal synthetic organelles via a simple biotin-avidin adhesion system. More advanced constructs are triggered externally by (i) light using protein-protein bindings or (ii) via the addition of ssDNA for DNA-DNA interactions. Finally, in an effort to reflect the complexity of eukaryotic cells we have designed, from the bottom-up, an enzymatic cascade pathway which takes full advantage of multiple membrane compartments with distinct reconstituted membrane pores. This synthetic signalling cascade is triggered by first capturing them in our microfluidic devices and second by using microfluidic flow to controllably deliver the chemical inputs.